Journal: Frontiers in Immunology

Article Title: Generation and characterization of a tamoxifen-inducible lineage tracing tool Cd2-P2A-CreERT2 knock-in mice

doi: 10.3389/fimmu.2025.1482070

Figure Lengend Snippet: Cre recombinase-expression and fluorescence signal in a Cd2-CreERT2 -inducible tdTomato mouse model. (A) Schematic diagram depicting the Cd2-CreERT2 mouse crossbred with a tdTomato reporter mouse line. (B) Schematic diagram depicting the Tamoxifen-induced Cre-recombinase. (C) Representative images of thymus, spleen and mesenteric lymph node (mLN) from Cd2-CreERT2;Rosa26-LSL-tdTomato mice visualized directly for tdTomato fluorescence. (D) Representative immunofluorescence images of tissue sections for tdTomato expression in CD2 + cells (green).

Article Snippet: The Cd2-CreERT2 mice were crossed with a loxP -flanked STOP cassette (LSL) Rosa26-LSL-tdTomato reporter line (AI9, Jackson Labs) ( ) to yield a Cd2 -driven tdTomato reporter mice ( Cd2-CreERT2;Rosa26-LSL-tdTomato ).The genotype of Cd2-CreERT2;Rosa26-LSL-tdTomato mice was identified using the same strategy as genotyping of Cd2-CreERT2 F2 generation for the Cd2 locus, and the protocol given by Jackson Labs for the Rosa26 locus.

Techniques: Expressing, Fluorescence, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: Generation and characterization of a tamoxifen-inducible lineage tracing tool Cd2-P2A-CreERT2 knock-in mice

doi: 10.3389/fimmu.2025.1482070

Figure Lengend Snippet: tdTomato + /CD2 + cells analysis from flow cytometry measurements. Representative flow cytometry plots of tdTomato + /CD2 + cells from CD45 + subpopulations of peripheral blood (PB), thymus, spleen (SP) and mesenteric lymph node (mLN) in Cd2-CreERT2;Rosa26-LSL-tdTomato female (A) or male (B) mice with or without tamoxifen treatment. (C) Quantification of tdTomato + /CD2 + % cells, n=4.

Article Snippet: The Cd2-CreERT2 mice were crossed with a loxP -flanked STOP cassette (LSL) Rosa26-LSL-tdTomato reporter line (AI9, Jackson Labs) ( ) to yield a Cd2 -driven tdTomato reporter mice ( Cd2-CreERT2;Rosa26-LSL-tdTomato ).The genotype of Cd2-CreERT2;Rosa26-LSL-tdTomato mice was identified using the same strategy as genotyping of Cd2-CreERT2 F2 generation for the Cd2 locus, and the protocol given by Jackson Labs for the Rosa26 locus.

Techniques: Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Generation and characterization of a tamoxifen-inducible lineage tracing tool Cd2-P2A-CreERT2 knock-in mice

doi: 10.3389/fimmu.2025.1482070

Figure Lengend Snippet: CD2-Cre activity in mature leukocytes (T cells). Co-expression of CD2 and tdTomato in T cells from peripheral blood (PB), thymus, spleen (SP) and mesenteric lymph node (mLN) gated on CD3 + cells from Cd2-CreERT2;Rosa26-LSL-tdTomato female (A) or male (B) mice with or without tamoxifen treatment (A, B) and graph bar (C) . (D–F) gated on CD4 + cells from Cd2-CreERT2; Rosa26-LSL-tdTomato female (A) or male (B) mice with or without tamoxifen treatment (D, E) and graph bar (F) , (G–I) Gated on CD8 + cells from Cd2-CreERT2;Rosa26-LSL-tdTomato female (A) or male (B) mice, with or without tamoxifen treatment (G, H) and graph bar (I) . (J) Co-expression of CD2 and tdTomato in CD4 + CD8 + double-positive (DP) T cells from the thymus of Cd2-CreERT2;Rosa26-LSL-tdTomato female or male mice with or without tamoxifen treatment. n=4.

Article Snippet: The Cd2-CreERT2 mice were crossed with a loxP -flanked STOP cassette (LSL) Rosa26-LSL-tdTomato reporter line (AI9, Jackson Labs) ( ) to yield a Cd2 -driven tdTomato reporter mice ( Cd2-CreERT2;Rosa26-LSL-tdTomato ).The genotype of Cd2-CreERT2;Rosa26-LSL-tdTomato mice was identified using the same strategy as genotyping of Cd2-CreERT2 F2 generation for the Cd2 locus, and the protocol given by Jackson Labs for the Rosa26 locus.

Techniques: Activity Assay, Expressing

Journal: Frontiers in Immunology

Article Title: Generation and characterization of a tamoxifen-inducible lineage tracing tool Cd2-P2A-CreERT2 knock-in mice

doi: 10.3389/fimmu.2025.1482070

Figure Lengend Snippet: tdTomato signal in B leukocyte populations. Numbers depict the percent of CD2 + tdTomato + cells in Cd2-CreERT2; Rosa26-LSL-tdTomato female (A) and male mice (B) in peripheral blood (PB), Spleen (SP) and mesenteric lymph node (mLN) gated on CD19 + and the graph bar (C) . (D–H) Gated on CD19 + IgD - (D, E) , CD19 + IgD + (F, G) and graph bar (H) n=4.

Article Snippet: The Cd2-CreERT2 mice were crossed with a loxP -flanked STOP cassette (LSL) Rosa26-LSL-tdTomato reporter line (AI9, Jackson Labs) ( ) to yield a Cd2 -driven tdTomato reporter mice ( Cd2-CreERT2;Rosa26-LSL-tdTomato ).The genotype of Cd2-CreERT2;Rosa26-LSL-tdTomato mice was identified using the same strategy as genotyping of Cd2-CreERT2 F2 generation for the Cd2 locus, and the protocol given by Jackson Labs for the Rosa26 locus.

Techniques:

Journal: Frontiers in Immunology

Article Title: Generation and characterization of a tamoxifen-inducible lineage tracing tool Cd2-P2A-CreERT2 knock-in mice

doi: 10.3389/fimmu.2025.1482070

Figure Lengend Snippet: tdTomato signal in NK leukocyte populations. Numbers depict the percent of CD2 + tdTomato + cells in Cd2-CreERT2; Rosa26-LSL-tdTomato female (A) and male mice (B) in peripheral blood (PB), Spleen (SP) and mesenteric lymph node (mLN) gated on NK1.1 + CD3 - and the graph bar (C) n=4.

Article Snippet: The Cd2-CreERT2 mice were crossed with a loxP -flanked STOP cassette (LSL) Rosa26-LSL-tdTomato reporter line (AI9, Jackson Labs) ( ) to yield a Cd2 -driven tdTomato reporter mice ( Cd2-CreERT2;Rosa26-LSL-tdTomato ).The genotype of Cd2-CreERT2;Rosa26-LSL-tdTomato mice was identified using the same strategy as genotyping of Cd2-CreERT2 F2 generation for the Cd2 locus, and the protocol given by Jackson Labs for the Rosa26 locus.

Techniques:

Journal: Frontiers in Immunology

Article Title: Generation and characterization of a tamoxifen-inducible lineage tracing tool Cd2-P2A-CreERT2 knock-in mice

doi: 10.3389/fimmu.2025.1482070

Figure Lengend Snippet: tdTomato expression in the indicated myeloid populations from Cd2-CreERT2;Rosa26-LSL-tdTomato mice. Flow cytometric analysis of tdTomato expression in macrophages (A) , granulocytes (B) monocytes (C) and dendritic cells (D) lineages. (E) The proportion of tdTomato + /CD2 + was charted as mean ± SEM, n = 4.

Article Snippet: The Cd2-CreERT2 mice were crossed with a loxP -flanked STOP cassette (LSL) Rosa26-LSL-tdTomato reporter line (AI9, Jackson Labs) ( ) to yield a Cd2 -driven tdTomato reporter mice ( Cd2-CreERT2;Rosa26-LSL-tdTomato ).The genotype of Cd2-CreERT2;Rosa26-LSL-tdTomato mice was identified using the same strategy as genotyping of Cd2-CreERT2 F2 generation for the Cd2 locus, and the protocol given by Jackson Labs for the Rosa26 locus.

Techniques: Expressing

Journal: JCI Insight

Article Title: Map2k6 is a potent genetic modifier of arterial rupture in vascular Ehlers-Danlos syndrome mice

doi: 10.1172/jci.insight.187315

Figure Lengend Snippet: Kaplan-Meier survival curve comparing ( A ) male 129 vEDS Map2k6 +/+ ( n = 17), vEDS Map2k6 +/– ( n = 20), and vEDS Map2k6 –/– mice ( n = 10) and ( B ) female 129 vEDS Map2k6 +/+ ( n = 19), vEDS Map2k6 +/– ( n = 15), and vEDS Map2k6 –/– mice ( n = 12). Significant differences were calculated using log-rank (Mantel-Cox) analysis (* P < 0.05; ** P < 0.01). ( C ) Immunoblot analysis of phosphorylated PKC at residue Ser 660 (pPKC) and phosphorylated ERK (pERK1/2) comparing aortic lysates obtained from the proximal descending aortas of mice at 2 months of age. ( D ) Quantification of p-PKC and p-ERK normalized to β-actin of control Map2k6 +/+ ( n = 3), vEDS Map2k6 +/+ ( n = 4), control Map2k6 –/– ( n = 7), and vEDS Map2k6 –/– ( n = 5) mice. P value refers to 2-way ANOVA with Holm-Šídák post hoc test (* P < 0.05, *** P < 0.001). ( E ) Immunofluorescence of sections from the proximal descending thoracic aorta of vEDS Map2k6 +/+ and vEDS Map2k6 –/– mice. The dashed line marks the approximate boundaries of the aortic wall. Scale bar is 50 microns. ( F ) Mean and total protein phosphatase 1 (PP1) dephosphorylation activity in aortic protein lysates from 129 vEDS Map2k6 +/+ mice ( n = 6) and 129 vEDS Map2k6 –/– ( n = 7). P value refers to unpaired t test with Welch’s correction (* P < 0.05, ** P < 0.01).DiFMU, 6,8-difluoro-7-hydroxy-4-methylcoumarin. In D and F , each symbol represents an independent biological replicate, with unfilled symbols representing male samples. Error bars show mean ± SEM. ( G ) Kaplan-Meier survival curve comparing control 129 vEDS Map2k6 –/– mice ( n = 33, 17 females and 16 males) with 129 vEDS Map2k6 –/– ( n = 11, 8 females and 3 males) mice receiving ruboxistaurin (PKC inhibitor) starting at postnatal day 21. Significant differences were calculated using log-rank (Mantel-Cox) analysis (* P < 0.05).

Article Snippet: Map2k6 –/– mice were genotyped according to The Jackson Laboratory protocols.

Techniques: Western Blot, Residue, Control, Immunofluorescence, De-Phosphorylation Assay, Activity Assay